In:
Clinical Chemistry and Laboratory Medicine (CCLM), Walter de Gruyter GmbH, Vol. 50, No. 2 ( 2012-01-1)
Abstract:
In acute myeloid leukemia (AML), the internal tandem duplication (ITD) in the juxtamembrane domain of the A complex evaluation of the analytical properties of the three most frequently used detection methods – PCR followed by agarose (AGE), polyacrylamide (PAGE) or capillary electrophoresis (CE) – was performed on 95 DNA samples obtained from 73 AML patients. All the three methods verified the presence of a mutant allele in 20 samples from 18 patients. AGE and PAGE could detect the presence of 1%–2% mutant allele, while the detection limit of CE was 0.28%. However, acceptable reproducibility (inter-assay CV 〈 25%) of the mutant allele rate determination was only achievable above 1.5% mutant/total allele rate. The reproducibility of the ITD size determination by CE was much better, but the ITD size calculated by PeakScanner or GeneScan analysis was 7% lower as compared to values obtained by DNA sequencing. The presence of multiple ITD was over-estimated by PAGE and AGE due to the formation of heteroduplexes. This study suggests the use of PCR+CE in the diagnostics and the follow-up of AML patients. The data further supports the importance of proper analytical evaluation of home-made molecular biological diagnostic tests.
Type of Medium:
Online Resource
ISSN:
1437-4331
,
1434-6621
DOI:
10.1515/cclm.2011.762
Language:
Unknown
Publisher:
Walter de Gruyter GmbH
Publication Date:
2012
detail.hit.zdb_id:
1492732-9
SSG:
15,3